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Investigation On The Application Of Phthalocyanine Compounds In Analytical Chemistry

Posted on:2002-09-02Degree:DoctorType:Dissertation
Country:ChinaCandidate:X L ChenFull Text:PDF
GTID:1101360062475573Subject:Analytical Chemistry
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Phthalocyanines are a category of compounds which are similar to that of porphyrins and chlorophyll in structure. Due to their ease to synthesize and good stability to light and heat, phthalocyanines have been widely applied in dyes, pigments, photoelectric materials, catalysts, photodynaniic therapy and fluorescence analysis. On the basis of forefathers' research, this work is going to exploit the application of phthalocyanines in fluorescence assay. Meanwhile, the application of the resonance light scattering of metallophthalocyanine compounds to the determination of biomacromolecules will also be explored. This dissertation is composed of five chapters.In chapter 1, the synthesis, structure, properties and applications of phthalocyanine compounds are briefly summarized. Emphasis is paid on their applications in fluorescence assay. Considering some areas which will be involved with in the work, brief reviews are also made on the following aspects: developments of fluorogenic substrates for H7RP and HRP mimics, and the application of reversed micelles in HRP-catalyzed fluorogenic reactionIn chapter 2, the feasibility of applying tetra-substituted amino aluminum phthalocyanine (TAA1Pc) as a new red-region fluorogenic substrate for HRP and HRP mimics is studied. The catalytic effect of HRP in the fluorogenic reaction between H,02 and TAA1Pc is first investigated. Spectral characteristics of the measurement system and catalytic parameters of HRP obtained from the Lineweaver-Burk plot indicate that LIRP exhibits good catalytic activity, which implies the feasibility of using TAMPc as a substrate for IIRP in fluorescence assay. Then, two novel sensitive fluorinietric methods are developed for the determination of trace 11202 based on using TAAIPc as a substrate for I-IRP and ILRP mimics. The proposed methods have advantages of high sensitivity and low detection limit. Moreover, they can effectively avoid the interference from the background fluorescence of biological samples since red-region excitation and emission wavelengths are employed (610 / 678 nm). These methods have been applied successfully to the determination of glucose in human sera and 11202 in rainwater with satisfactory results. Finally, the study is concentrated on applying the proposed LIRP- or HRP mimic-enzymatic reaction system with TAA1Pc as a fluorogenic substrate to develop some enzyme-labeled fluoroimmunoassays including competitive iminunoassay for c~-AFP and sandwich immunoassay for flBsAg. In the competitive immunoassay, monoclonalanti-AFP antibody is coated on a 96-well plate (polystyrene). Non-labeled and HRPlabeled APP compete for binding to the plate-bound antibody. After the immunoreaction, the immunochemically adsorbed HRP-AFP conjugate moiety is' determined by measuring the fluorescence intensity of the HRP-AFP-TAA1Pc-H202 system. The method has been applied to determine the AFP level in colon carcinoma patient sera and the results are satisfactory. In the sandwich immunoassay, standard HBsAg or the sample to be determined reacts first with anti-HBsAg antibody coated on a 96-well plate, then further reacts with the fixed amount of HRP-labeled anti-HBsAg. After the two-step immunoreaction, the immunochemically adsorbed HRP-anti-HBsAg conjugate moiety is determined by measuring the fluorescence intensity of the HRP-anti-HbsAg-TAA1Pc-H202 system. The feasibility of this method for determining the HBsAg level in healthy human serum and hepatitis B patient serum is further examined and the results demonstrate that TAAIPc is a promising substrate for HRP-labeled fluoroimmunoassays and offers good prospects in biochemical analysis.In chapter 3, the catalytic behavior of iron tetrasulfonatophthalocyanine (FeTSPc) in reversed micelles is investigated for the oxidation reaction of L-tyrosine with H202. A reversed micelle is a surfactant aggregate formed in an organic solvent that usually contains a small amount of water. Due to its unique solubility, self-assembly and relative organized structures, reversed micelles can we...
Keywords/Search Tags:Phthalocyanine compounds, HRP-enzymatic reaction, reversed micelles, ion-association, resonance light scattering, biomacromolecules.
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